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《Acta Agrestia Sinica》 2001-02
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Optimization of Alfalfa Genomic DNA Extraction And RAPD Conditions

YUAN Qing hua 1,GUI Zhi 2,ZHANG Wen shu 1 (1.Institute of Animal Science,CAAS,Beijing 100094,China;2.Gansu Agriculture University,Lanzhou 730070,China)  
Four different methods were tested in isolation of alfalfa (Medicago sativa) genomic DNAs.The results show that the reformed CTAB is more effective.Based on the genomic DNA extracted from alfalfa,essential factors affecting the result of RAPD assay were tested and compared.An optimal reaction system suitable for the assay and usage of RAPD in alfalfa was established.This optimal of 25 μL reaction solution contained 2.5 μL 10x buffer,2.5 mmol/L MgCl 2,0.3 mmol/L each dNTP,0.1 μmol/L random primer,174 ng/μL template DNA,1 U Taq polymerase.The reaction program was devised for one cycle at 94℃ 4 minutes,36℃ 30 seconds,72℃ 1 minute,which is followed by 45 cycles,each with 30 seconds at 94℃,30 seconds at 36℃,1 minutes at 72℃,and a final extension at 72℃ for 10 minutes.
【Fund】: 国家自然科学基金! (编号 39870 5 5 9)资助项目
【CateGory Index】: Q943
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